Cancer stem cells from colorectal cancer-derived cell lines

Abstract

Cancer stem cells (CSCs) are the subpopulation of cells within a tumor that can self-renew, differentiate into multiple lineages, and drive tumor growth. Here we describe a two-pronged approach for the identification and characterization of CSCs from colorectal cancer cell lines, using a Matrigel-based differentiation assay, and cell surface markers CD44 and CD24. About 20 to 30% of cells from the SW1222 cell line form megacolonies in Matrigel that have complex 3D structures resembling colonic crypts. The megacolonies' capacity to self-renew in vitro is direct evidence that they contain the CSCs. Furthermore, just 200 cells from SW1222 megacolonies initiate tumors in NOD/SCID mice. We also showed that CD44 + CD24 + cells enriched for colorectal CSCs in the HT29 and SW1222 cell lines, which can self-renew and reform all four CD44/CD24 subpopulations, are the most clonogenic in vitro and can initiate tumors in vivo. A single SW1222 CD44 + CD24 + CSC, when grown in Matrigel, can form large megacolonies that differentiate into enterocyte, enteroendocrine, and goblet cell lineages. The HCT116 line does not differentiate or express CDX1, nor does it contain subpopulations of cells with greater tumor-forming capacity, suggesting that HCT116 contains mainly CSCs. However, forced expression of CDX1 in HCT116 leads to reduced clonogenicity and production of differentiating crypt-containing colonies, which can explain the selection for reduced CDX1 expression in many colorectal cancers. In summary, colorectal cancer cell lines contain subpopulations of CSCs, characterized by their cell surface markers and colony morphology, which can self-renew and differentiate into multiple lineages.clonogenicity | matrigel | CD44 | goblet cells

Metformin and soybean-derived bioactive molecules attenuate the expansion of stem cell-like epithelial subpopulation and confer apoptotic sensitivity in human colon cancer cells

“…In these studies, we primarily utilized the human colon cancer cell line HCT116 since its highly aggressive nature and increased proportion of non-differentiating CSCs are clinically correlated with lymph node metastases and reduced survival (Yeung et al 2010). We show that relative to the more differentiated, less aggressive colon cancer cell line HT29, HCT116 cells exhibit a greater proportion of the CSC subpopulation (designated CD133 ?…”

“…The human colon cancer cell lines HT29 and HCT116 contain a sub-population of cancer stem cell-like cells (CSC), which are distinguished by their expression of cell surface antigens CD133 and CD44 (Sahlberg et al 2014) and their ability to initiate tumors in nude mice (Yeung et al 2010;Chen et al 2011). The relative abundance of the CD133 ?…”

“…Here, we evaluated the effects of MET and several bioavailable and bioactive molecules present in soy protein isolate, namely the isoflavone Gen, peptide lunasin (Lun), and proteins b-conglycinin (b-con) and glycinin (Gly), alone and in combination, on the survival of CSCs present in the human colon cancer cell line HCT116 (Yeung et al 2010). We examined potential chemo-preventive mechanisms of these components in CSCs and determined whether the combination of MET, Gen, and Lun with 5-FU could increase CRC cell death relative to 5-FU alone.…”

Multiple uses of basement membrane‐like matrix (BME/Matrigel) in vitro and in vivo with cancer cells

“…Stem cells form megacolonies that can self-renew, differentiate, and form tumors in vivo. 15 The subpopulation of cells that form small colonies after passage, do not form tumors in mice. This simple approach can be used to select cells within a cell line of differing malignant potential.…”

“…14 Other assays based on cancer cell morphology, clonogenicity, growth, and dormancy on the matrix have also been developed more recently. 15,16 Coinjection of the basement membrane-like matrix with cancer cells either subcutaneously or in orthotopic sites was found to increase both the 'take' and growth of tumors in mice. [17][18][19] Thus, cancer cell interactions with the basement membrane have multiple uses both in vitro and in vivo (Table 2).…”